Whitening and antionxidative cosmetic composition containing resveratrol and method for preparing the same

ABSTRACT

Provided are a stabilized, skin-whitening and antioxidative cosmetic composition including resveratrol as a main component and either a primary stabilizer selected from cyclodextrin, polyethyleneglycol, and a mixture thereof, or a combination of the primary stabilizer and a secondary stabilizer selected from an alpha-lipoic acid, a water-soluble whitening composition, a phellodendron extract, an alteromonas ferment extract, and a mixture thereof, and a method for preparing the same. The cosmetic composition provides excellent skin whitening and antioxidative effects for a long-term even when used in a small quantity. Furthermore, solubility of the resveratrol and the alpha-lipoic acid in polyol or ethanol is enhanced, thereby improving low use satisfaction and skin irritation that may be caused due to excess use of an organic solvent. In addition, problems such as crystal precipitation at low temperature, off-flavor and discoloration upon exposure to sun light can be solved.

TECHNICAL FIELD

The present invention relates to a whitening and antioxidative cosmeticcomposition containing resveratrol and a method for preparing the same.More particularly, the present invention relates to a cosmeticcomposition containing resveratrol as a main component and either aprimary stabilizer selected from cyclodextrin, polyethyleneglycol, and amixture thereof, or a combination of the primary stabilizer and asecondary stabilizer selected from an alpha-lipoic acid, a water-solublewhitening composition, a phellodendron extract, an alteromonas fermentextract, and a mixture thereof, and a method for preparing the same.

BACKGROUND ART

The skin is a very important organ that protects the internal organs ofthe human body and effects biochemical and physical functions whilebeing in direct contact with external environment. It is largelycomposed of three different layers, i.e., epidermis, dermis, andsubcutaneous tissue. Skin color in humans is mainly determined bynumber, size, type, and distribution of melanosomes containing melaninthat are spread throughout skin cells. Melanosomes are produced bymelanin cells. Melanin is a dark pigment which is produced inmelanocytes of epidermis.

Melanin absorbs the energy of UV light irradiated by sun light andprevents the light from penetrating into deeper skin cells. However,when melanin production is abnormally low, a skin disease such asleukoderma may be caused. On the other hand, overproduction of melaninby UV exposure and the like may cause a skin damage such as freckles andlentigo-type freckles and may contribute to development of a skincancer.

Resveratrol is present in a wide variety of gymnosperms and angiospermsand is mainly synthesized in the form of glycoside linked with sugargroup even though some of resveratrol is in free state [Gorham, J.,Prog. Phytochem., 6:203-209, 1980]. Resveratrol exists in cis and transisoforms. Stable trans-resveratrol is common in nature [Trela, B. C. &Waterhouse, A, L., J. Agric. Food Chem., 44(5): 1253-1257, 1996].

Resveratrol is mainly found in the shell, seed, stem, and leaf ofgrapes. In particular, resveratrol is abundantly contained in grapessuch as Vitis vinifeera, Vitis rotundifolia, and Vitis labrusca.Resveratrol is also found in wines made from these grapes, and extractsand powders of the aforementioned natural sources. In addition,resveratrolis contained in roots of Polygonum sp. such as Polygonumcuspidatum and Polygonum multiflorum, belonging to the Polygonaceaefamily.

As used herein, the term “resveratrol” coverstrans-3,4,5-trihydroxystilbene (i.e., suitable resveratrol) asrepresented by the following Formula 1 and its cis-isomers, glycosidesthereof, and resveratrol-containing extracts and powders obtained from asuitable plant.

Trans-resveratrol has been reported as a physiologically activesubstance that has an anticancer effect (Jang M., et al., Sci.,275:218-220, 1997), antithrombotic effect (Chung, M. L. et al., PlantaMed., 58:274-276, 1992; Frankel, E. N. et al., Lancet, 341:1103-1104,1993; Bertrilli, A. A. E. et al., Int. J. Tissue React., 17:1-3, 1995;Pae-Asciak, C. R. et al., Clin. Chim. Acta., 235:207-219, 1995), antiinflammatory effect (Kimura, Y. et al., Biochim. Biophys. Acta.,834:275-278, 1985), and antihyperlipidemic effect (Arichi, H., et al,chem. Pharm. Bull., 30(5):1766-1770, 1982).

In particular, recent studies have shown that trans-resveratrol is aphytoestrogen that produces estrogen-like effects, and thus, there hasbeen an increasing interest in trans-resveratrol as phytoestrogen (Gehm,B. D. et al., Proc. Nasl. Acad. Sci., 94:14138-14143, 1997). A cosmeticcomposition containing a grape extract is disclosed in Japanese PatentNo. 06336421, U.S. Pat. Nos. 5,683,683, 5,439,672, and 5,171,577.PCT/EP98/04223 discloses a cosmetic composition containing resveratrol.According to the disclosure in PCT/EP98/04223, the cosmetic compositionprevents proliferation and differentiation of keratinocytes or skinirritation by alpha-hydroxy acid. However, this patent document issilent about solutions to problems such as low use satisfaction, skinirritation, and crystal precipitation at low temperature that may becaused by excess use of polyol or ethanol that is used as a solvent in acosmetic product due to low solubility of resveratrol in the polyol orthe ethanol, and off-flavor and discoloration upon exposure to sunlight.

In spite of excellent whitening and antioxidative effects, there is aproblem in that resveratrol may be precipitated as a crystal when usedin a cosmetic composition containing water. For this reason, excessethanol or polyol must be used. That is, 0.1 wt % of resveratrolrequires use of 10 wt % of an organic solvent such as polyol, whichmakes it difficult to use a high content of resveratrol. A high contentof resveratrol can be used only in a substantially water-free cosmeticcomposition. However, a water-free cosmetic composition using only anorganic solvent such as ethanol or polyol is hardly commerciallyavailable due to severe skin irritation and low use satisfaction. Inparticular, vitamin C that is difficult to be stable in a cosmeticcomposition is prepared as a “polyol in silicone” or “silicone inpolyol” system. However, this formulation system has low usesatisfaction, complex preparation process, and expensive material cost,and thus, is limitedly used in a cosmetic product.

Meanwhile, alpha-lipoic acid as used herein has strong antioxidativeactivity. It is known that alpha-lipoic acid serves to facilitateproduction of glutathione that is another antioxidative substance of thehuman body or help functions and regeneration of vitamin E and C,thereby increasing an antioxidative effect in the human body.

Based on these effects of alpha-lipoic acid, alpha-lipoic acid iscurrently used in the treatment of diabetes, nervous disease, cataract,heart failure, atherosclerosis, and the like. In addition, alpha-lipoicacid is known to have excellent moisturizing capability by ceramideproduction (U.S. Pat. No. 5,472,698A), therapeutic effects of skininflammation, necrosis, tumor, allergy, and the like (Germany Patent No.441,708A), therapeutic effects of allergy, inflammation, and tanning (WO9508564A), skin whitening and tyrosinase inhibitory activity (JapanesePatent No. 63008315A), and acne prevention and relief by maintenance ofhomeostasis (Korean Patent Laid-Open Publication No. 1999-025848).However, there may arise problems such as low use satisfaction by excessuse of polyol in a cosmetic product due to low solubility ofalpha-lipoic acid in the polyol, unique odor due to a sulfur element,and off-flavor and discoloration due to separation of thiol group uponexposure to light.

Therefore, the present inventors have developed a whitening andantioxidative cosmetic composition which stably contains resveratrolthat had been difficult to be used in a cosmetic product, and thus, hasno side effects such as skin irritation.

In view of the above problems, the present invention provides a cosmeticcomposition containing resveratrol which is excellent in whitening andantioxidative effects and a method for preparing the same.

The above and other objects of the present invention can be accomplishedby embodiments of the present invention as will be describedhereinafter.

DISCLOSURE OF THE INVENTION

Therefore, according to an aspect of the present invention, there isprovided a whitening and antioxidative cosmetic composition includingresveratrol.

The resveratrol may be used in an amount of 0.001-10.0 wt %, based onthe total weight of the cosmetic composition.

The cosmetic composition may further include a primary stabilizerselected from the group consisting of cyclodextrin, polyethyleneglycol,and a mixture thereof.

The primary stabilizer may be used in an amount of 0.1 to 50 wt %, basedon the total weight of the cosmetic composition.

The cosmetic composition including the resveratrol and the primarystabilizer may further include a secondary stabilizer selected from thegroup consisting of an alpha lipoic acid, a water-soluble whiteningcomposition, a phellodendron extract, an alteromonas ferment extract,and a mixture thereof.

The secondary stabilizer may be used in an amount of 0.01 to15.0 wt %,basedonthe total weight of the cosmetic composition.

The water-soluble whitening composition may be selected from the groupconsisting of a citrus unshiu peel extract, an aminoethylphosphinicacid, Waltheria indica extract, Guava Phenone, an aqueous hydrolyzedyeast solution, and a mixture thereof.

According to another aspect of the present invention, there is provideda method for preparing a whitening and antioxidative cosmeticcomposition, including: (a) mixing a solvent and a primary stabilizer;(b) adding resveratrol to the mixture of step (a); and (c) adding asecondary stabilizer selected from the group consisting of analpha-lipoic acid, a water-soluble whitening composition, aphellodendron extract, an alteromonas ferment extract, and a mixturethereof, to the mixture of step (b).

Hereinafter, the present invention will be described in detail.

The present invention provides a whitening and antioxidative cosmeticcomposition. More particularly, the present invention provides acosmetic composition including a stabilized resveratrol and a primarystabilizer selected from cyclodextrin, polyethyleneglycol, and a mixturethereof. Preferably, the cyclodextrin is hydroxypropyl beta cyclodextrinand the polyethyleneglycol is polyethyleneglycol 400 orpolyethyleneglycol 32.

The cosmetic composition including the resveratrol and the primarystabilizer may further include a secondary stabilizer selected from thegroup consisting of an alpha-lipoic acid, a water-soluble whiteningcomposition, a phellodendron extract, an alteromonas ferment extract,and a mixture thereof.

The water-soluble whitening composition (Whitegenic, Korean PatentApplication No. 10-2002-0009503) may be selected from the groupconsisting of a citrus unshiu peel extract, an aminoethylphosphinicacid, Waltheria indica extract, Guava Phenone, an aqueous hydrolyzedyeast solution, and a mixture thereof.

In more detail, in the cosmetic composition including the stabilizedresveratrol, the resveratrol maybe used in an amount of 0.001-10.0 wt %,preferably 0.1-5.0 wt %, based on the total weight of the cosmeticcomposition. The primary stabilizer may be used in an amount of 0.1-50wt %, preferably 1.0-20.0 wt %, based on the total weight of thecosmetic composition. The secondary stabilizer may be used in an amountof 0.01-15.0 wt %, preferably 0.1-10.0 wt %, based on the total weightofthecosmetic composition.

In the cosmetic composition including the stabilized resveratrol, thebalance except the resveratrol, the primary stabilizer, and thesecondary stabilizer is a solvent selected from polyol, ethanol, andwater.

The alpha-lipoic acid with antioxidiative activity as used herein isrepresented by the following Formula 2. The alpha-lipoic acid is aco-factor that is essential in energy metabolism or cellular respirationof organism including microorganisms and humans, together with thioticacid, thioctan, and 1.2-dithiolane-3-pentanoic acid. Thealpha-lipoicacid has been recognized as a strong antioxidant since itwas first isolated in 1950.

The alpha-lipoic acid is soluble in ethanol or polyol which is generallyused in a cosmetic composition. However, the alpha-lipoic acid has adisadvantage in that it is quickly converted to dihydrolipoic acidrepresented by the following Formula 3, in a solution, therebygenerating an odor.

According to the cosmetic composition including the stabilizedresveratrol, the resveratrol is used in an amount of 0.001-10.0 wt %. Ifthe content of the resveratrol is less than 0.001 wt %, a desiredantioxidative effect may not be obtained. On the other hand, if thecontent of the resveratrol exceeds 10.0 wt %, an antioxidative effectrelative to an increased material cost may be insignificant.

A solvent as used herein may be a solvent commonly used in the cosmeticindustry, for example ethanol, 1,3-butyleneglycol, propyleneglycol,dipropyleneglycol, pentyleneglycol, or glycerine. Ethanol is irritatingand generates a specific smell when used in a high content. Therefore,it is preferable to use ethanol in an amount of about 10 wt % or less.In this regard, ethanol is not used in a cosmetic composition for asensitive skin. A cosmetic composition including a high content ofpolyol may cause sticky skin feel, thereby lowering use satisfaction.Therefore, it is preferable to use polyol in an amount of 10 wt % orless.

The phellodendron extract that is used herein as a stabilizer is anextract obtained by extracting Phellodendron plant with ethanol or1,3-butyleneglycol. The bark of Phellodendron amurense Ruprecht orRutaceae is used. The phellodendron plant is cold, bitter, andnon-toxic. It is effective in treatment of fever and jaundice that areconcentrated in the five viscera and the stomach and intestines,diarrhea and dysentery, gynecopathy, scabies and scabs, eye fever andache, sore throat, and a high fever that can melt bone. In addition, itis well known that the phellodendron plant has antifungal,antiinflammatory, and antivirus effects. However, there have been noreports that the phellodendron plant can prevent discoloration andoff-flavor by exposure to UV light. The present inventors found that thephellodendron plant can prevent phase instability of effectivecomponents by sun light and the like. The phellodendron extract iscommercially available under the trade name of Phellodendron Bark BG(Koei, Japan).

The alteromonas ferment extract that is used herein as anotherstabilizer is deepsane composed of 11 glucosides that is a fermentationproduction of Alteromonas macleodii that is a microorganism present in asevere environment having seawater depth of 3,000 m or atmosphericpressure of 300 bar. The present inventors found that the alteromonasferment extract has a molecular weight of 1.8×10⁶ Dalton and a skinirritation relief effect. The alteromonas ferment extract iscommercially available under the trade name of Abyssine 657 (Lanatech,France).

In the present invention, the secondary stabilizer selected from thewater-soluble whitening composition, the alpha-lipoic acid, thephellodendron extract, the alteromonas ferment extract, and the mixturethereof may be used in an amount of 0.01-15 wt %, based on the totalweight of the cosmetic composition. If the content of the secondarystabilizer exceeds 15 wt %, an ef fect enhancement relative to anincreased material cost may be insignificant.

The cyclodextrin and the polyethyleneglycol used as the primarystabilizer in the cosmetic composition including the resveratrol may berepresented by the following Formulae 4a and 4b, respectively.Preferably, the cyclodextrin is hydroxypropyl beta cyclodextrin and thepolyethyleneglycol is polyethyleneglycol 400 or polyethyleneglycol 32.

 H(OCH₂CH₂)nOHn=32, 400   <Formula 4b>

The primary stabilizer must be used in an amount of 5-20 wt %, based onthe total weight of effective components or in an amount of 0.1-50 wt %,based on the total weight of the cosmetic composition including thestabilized resveratrol. If the content of the primary stabilizer exceeds20 wt %, based on the total weight of the effective components, a costincrease and use satisfaction lowering by excess use of the primarystabilizer may be caused.

The cosmetic composition including the stabilized resveratrol of thepresent invention can solve phase instability with time such asdiscoloration, off-flavor, and crystal precipitation that may be causedduring preparation or storage and reduce skin irrigation, simultaneouslywith exhibiting continuously an excellent antioxidative activity.

A method for preparing the cosmetic composition including the stabilizedresveratrol according to the present invention includes:

(a) mixing a solvent and a primary stabilizer;

(b) adding the resveratrol to the mixture of step (a); and

(c) adding a secondary stabilizer selected from a water-solublewhitening composition, an alpha-lipoic acid, a phellodendron extract, analteromonas extract, and a mixture thereof, to the mixture of step (b).

In the method for preparing the cosmetic composition including theresveratrol, the solvent may be polyol, ethanol, or water.

In step (a), the primary stabilizer may be cyclodextrin orpolyethyleneglycol, and preferably, hydroxypropyl beta cyclodextrin,polyethyleneglycol 32, polyethyleneglycol 400, or a mixture thereof.

The method for preparing the cosmetic composition including theresveratrol is generally carried out at room temperature and underatmospheric pressure. If necessary, heating may be used.

In the cosmetic composition including the resveratrol, the primarystabilizer is used in an amount of 1.0-50.0 wt %, based on the totalweight of the cosmetic composition.

A cosmetic composition of the present invention may be formulated as allformulations that can be prepared by solubilization, emulsification, ordispersion, for example toner, lotion, cream, essence, sunscreen cream,cleansing foam, cleansing cream, or pack, but are not limited thereto.

BEST MODE FOR CARRYING OUT THE INVENTION

To solve problems such as low solubility of resveratrol that restrictsits utility in a cosmetic composition in spite of excellentantioxidative effect and the like, and discoloration, off-flavor, andcrystal precipitation during long-term storage, a cosmetic compositionof the present invention includes resveratrol stabilized by a primarystabilizer or a combination of the primary stabilizer and a secondarystabilizer. In Experimental Examples as will be described hereinafter,to evaluate long-term effects with time of the cosmetic composition, along-term effect of an antioxidative activity is evaluated by measuringradical scavenging ability. In addition, skin safety is evaluated byskin irritation test based on patch test in skins of healthy adults.Phase stability in the cosmetic composition is evaluated by observingcolor change and crystal precipitation with time after exposure to 40°C., 4° C., and sun light.

The whitening activity of the cosmetic composition including theresveratrol is evaluated by measuring inhibitory activity againsttyrosinase that contributes to melanin biosynthesis, radical scavengingactivity, and antiproliferative activity against melanoma cells thathave a similar condition to living system.

According to these test results, the cosmetic composition including theresveratrol and either the primary stabilizer or a combination of theprimary stabilizer and the secondary stabilizer of the present inventionexhibits an excellent whitening activity, phase stability, and skinsafety, relative to a commercially available whitening agent such asalbutin and cojic acid, and thus, can be efficiently used as a whiteningcosmetic composition.

Hereinafter, the present invention will be described more specificallyby Examples but the present invention is not limited to or by them.

EXPERIMENTAL EXAMPLE 1 Tyrosinase Inhibitory Activity Test

Inhibitory activity of resveratrol against tyrosinase that participatesin main processes of melanin biosynthesis was measured to evaluate awhitening effect of resveratrol and the result is presented in Table 1below.

Tyrosine used as a substrate in main processes of melanin biosynthesis,mushroom tyrosinase, or tyrosinase fragments separated from melanocyteswas added to test samples and incubated for a predetermined time. Theabsorbance of dopachrome that was a reaction product was measured at anabsorption wavelength to evaluate the whitening effects of the testsamples.Tyrosinase inhibitory activity (%)=100−((OD _(exp) −OD _(con))/OD_(std)×100)

OD_(exp): Absorbance of test sample containing tyrosinase

OD_(con): Absorbance of test sample containing no tyrosinase

OD_(std): Absorbance of standard sample containing tyrosinase TABLE 1Test results of tyrosinase inhibitory activity Sample *IC₅₀ (μg/ml)Resveratrol 2 Cojic acid 5 Available licorice extract >50 White mulberrypowder >50 Vitamin C >100 Albutin >100 Water-soluble whitening >100composition (*Whitegenic) White mulberry extract >1000 Grapeseed extract>1000*IC₅₀: 50% tyrosinase activity inhibition concentration (as the value ofIC₅₀ decreases, tyrosinase inhibitory activity increases)*Whitegentic (trade name, Korea):a mixture of Whitegentic 1 (Sederma,France):Whitegenic 2 (Exymol, Monaco):Whitegenic 3 (Sero, France) orGuava Phenone (10% aqueous solution):Whitegenic 4 (Sapporo, Japan)(5:1:0.3:2)

It can be seen from Table 1 that the resveratrol and the cojic acid haveexcellent tyrosinase inhibitory activity.

EXPERIMENTAL EXAMPLE 2 Free Radical Scavenging Activity Test

To evaluate the antioxidative activity of the resveratrol, a freeradical scavenging activity test was performed as the follows and theresults are presented in Table 2 below.

When methanol solutions of test samples were converted to clearsolutions due to scavenging of relatively stable deep blue-coloredDPPH(1,1-diphenyl-2-hydrazyl) radicals, the degree of reduction of theabsorbance of the methanol solutions was measured at 516 nm.Free radical scavenging activity (%)=100−( (OD _(exp) −OD _(con))/OD_(std)×100)

OD_(exp): Absorbance of test sample containing DPPH

OD_(con): Absorbance of test sample containing no DPPH

OD_(std): Absorbance of standard sample containing DPPH TABLE 2 Testresults of free radical scavenging activity Sample IC₅₀ (μg/ml) VitaminC 3 Resveratrol 10 Albutin >50 White mulberry powder >100 Cojicacid >200 Available licorice extract >250 Grapeseed extract >250Water-soluble whitening composition >250 (*Whitegenic) White mulberryextract >1000*IC₅₀: 50% free radical scavenging concentration (as the value of IC₅₀decreases, free radical scavenging activity increases)*Whitegenic (trade name, Korea): a mixture of Whitegenic 1 (Sederma,France):Whitegenic 2 (Exymol, Monaco):Whitegenic 3 (Sero, France) orGuava Phenone (10% aqueous solution):Whitegenic 4 (Sapporo, Japan)(5:1:0.3:2)

It can be seen from Table 2 that Vitamin C and resveratrol are effectivefree radical scavengers.

EXPERIMENTAL EXAMPLE 3 Whitening Activity Test Based on Proliferation ofSkin Melanocytes

Inhibitory activity of resveratrol against melanin production inmelanocytes of healthy persons was measured and the results arepresented in Table 3 below.

The whitening activity test based on proliferation of skin melanocyteswas performed as the following method:

-   -   1. Human skin melanocytes in MGM (Melanocyte Growth Media,        Clonetics) media were incubated at a 5% CO₂ incubator at 37° C.    -   2. A predetermined number of cells (2×10⁶) were cultured in75T        culture flasks. When cell confluence was identified, test        samples of a predetermined concentration were loaded in the cell        cultures and incubated for 2-3 days.    -   3. The cultures were removed, washed with phosphate buffered        saline (PBS), and treated with trypsin solution, to harvest        cells.    -   4. The harvested cells were centrifuged to obtain cell pellets.

The colors of the cell pellets were observed. The cell pellets weredissolved in 1N NaOH and the absorbance was measured at 475 nm.

-   -   5. The concentration of melanin was calculated by using        absorbance values obtained from melanin standard and then        converted to the concentration of melanin/the total protein        concentration of melanocytes.        Antiproliferative activity against melanoma        cells(%)=[(A−B)/A]×100

A: concentration of melain/concentration of protein in control

(control: melanocytes normally grown in a culture medium containing notest samples)

B: concentration of melain/concentration of protein in each test sampleTABLE 3 Test results of melanin production inhibitory activity(concentration of each test sample: 0.02%) Melanin production inhibitorySample activity (%) Resveratrol 53.17 Water-soluble whitening 35.00composition (*Whitegenic) Cojic acid 22.62 Vitamin C 19.72 Availablelicorice extract 13.20 White mulberry powder 5.1 Albutin 2.00 Whitemulberry extract 1.80 Grapeseed extract 1.52*Whitegenic (trade name, Korea): a mixture of Whitegenic 1 (Sederma,France):Whitegenic 2 (Exymol, Monaco):Whitegenic 3 (Sero, France) orGuava Phenone (10% aqueous solution):Whitegenic 4 (Sapporo, Japan) =5:1:0.3:2

It can be seen from Table 3 that the resveratrol and the water-solublewhitening composition (Whitegenic, Korea) exhibit excellent inhibitoryactivity of melanin production.

Based on the test results of Experimental Examples, cosmeticcompositions capable of sustaining and enhancing the whitening effect ofthe resveratrol were prepared. To evaluate whether skin whiteningactivity can be maintained while maintaining excellent skin safety andphase stability, the whitening effect tests of the cosmetic compositionswere performed according to the aforementioned melanocytes proliferationtest.

EXAMPLES 1-7 AND COMPARATIVE EXAMPLES 1-8 Preparation of HighlyConcentrated and Stabilized Cosmetic Compositions

To evaluate an enhancement effect of solubility and phase stability ofresveratrol, stabilized cosmetic compositions were prepared according tocomposition ratios presented in Table 4 below (Table 4a for Examples1-7, Table 4b for Comparative Examples 1-8). Each composition ofExamples 1-7 and comparative Examples 1-8 was prepared as the follows.

First, a solvent part including primary stabilizers was sufficientlystirred to obtain a clear solution. Resveratrol used as an effectivecomponent was gradually added to the clear solution at room temperatureand stirred for 30-60 minutes. After the resveratrol was completelydissolved, components of a secondary stabilizer part were graduallyadded thereto and stirred for 10-20 minutes to there by obtain astabilized cosmetic composition. TABLE 4a Stabilized cosmeticcomposition Section Component Exam. 1 Exam. 2 Exam. 3 Exam. 4 Exam. 5Exam. 6 Exam. 7 Solvent Purified To 100 To 100 To 100 To 100 To 100 ToTo part water 100 100 Hydroxypropyl 10.0  20.0  30.0  50.0  50.0  — —beta cyclodextrin¹⁾ Polyethyleneglycol²⁾ — — — — — 20.0  —1,3-butyleneglycol — — — — — — 5.0 Ethanol — — — — — — 5.0 EffectiveResveratrol 0.5 — 2.0 5.0 5.0 2.0 0.5 component Secondary Waste-soluble— — — — — — 20.0  stabilizer whitening part composition Alpha-lipoic —2.0 3.0 5.0 5.0 5.0 — acid Phellodendron 1.0 3.0 5.0 10.0  10.0  10.0  —extract³⁾ Alteromonas — 1.0 3.0 5.0 — — — ferment extract⁴⁾Exam.: Example¹⁾Celdex HP-beta-CD (Food and Chemical Engineering, Ltd., JAPAN)²⁾Renex PEG 1500, PEG-400 (Uniqema Americas)³⁾Pellodendron Bark BG (Koei's, JAPAN)⁴⁾Abyssine 657 (Lanatech, FRANCE)

TABLE 4b Stabilized composition Section Component Comp. 1 Comp. 2 Comp.3 Comp. 4 Comp. 5 Comp. 6 Comp. 7 Comp. 8 Solvent Purified water To ToTo To To To To To part 100 100 100 100 100 100 100 100 Hydroxypropylbeta 3.0 10.0  — 50.0  — — — — cyclodextrin¹⁾ Polyethyleneglycol²⁾ — — —— — — — 1,3-butyleneglycol — — 30.0  — 50.0  5.0 — Ethanol — — 10.0  —20.0  15.0  10.0  10.0 Effective Resveratrol 1.0 2.0 3.0 5.0 5.0 0.5 — —component Secondary Water-soluble — — — — — — — 20.0 stabilizerwhitening part composition Alpha-lipoic acid — — 5.0 5.0 — — —Phellodendron — — 3.0 — — — — — extract³⁾ Alteromonas ferment — — 1.0 —— — — — extract⁴⁾Comp.: Comparative Example¹Celdex HP-beta-CD (Food and Chemical Engineering, Ltd., JAPAN)²⁾Renex PEG 1500, PEG-400 (Uniqema Americas)³⁾Phellodendron Bark BG (Koei's, JAPAN)⁴⁾Abyssine 657 (Lanatech, FRANCE)

EXPERIMENTAL EXAMPLE 4 Solubility and Phase Stability Test

To compare the solubility and phase stability of resveratrol between thecompositions of the present invention and conventional compositionscontaining a common solvent such as purified water, polyol and ethanol,solubility, flavor, and color were observed immediately afterpreparation and at predetermined times after preparation and the resultsare presented in Tables 5 and 6 below. TABLE 5 Solubility Immediatelyafter One month after Section preparation preparation Example 1Dissolution Good Example 2 Dissolution Good Example 3 Dissolution GoodExample 4 Dissolution Good Example 6 Comparative Example 1 Nodissolution Crystal precipitation (presence of crystal) ComparativeExample 2 Dissolution Good Comparative Example 3 Dissolution Crystalprecipitation Comparative Example 4 Dissolution Good Comparative Example5 Dissolution Crystal precipitation

As shown in Table 5, with respect to the compositions of Examples 1-4and 6 according to the present invention, even when the content of theeffective component (resveratrol) was 15 wt %, solubility was good. Evenat one month after preparation, crystal precipitation was not observed.As described above, the hydroxypropyl beta cyclodextrin must be used in5-20 wt %, based on the total weight of the effective component. Withrespect to the composition of Comparative Example 1 in which the contentof the hydroxypropyl beta cyclodextrin was less than 5 wt %, crystalswere present due to incomplete encapsulation. On the other hand, if thecontent of the hydroxypropyl beta cyclodextrin exceeds 20 wt %, anincrease of a material cost and lowering of use satisfaction due toexcess use of the encapsulating agent may be caused. Therefore, it canbe seen that use of a suitable amount of the encapsulating agent isrequired. TABLE 6 Phase stability Section 1 day 1 month 2 months 3months Example 1 R.T. ⊚ ⊚ ⊚ ⊚ 40° C. ⊚ ⊚ ⊚ 0  4° C. ⊚ ⊚ ⊚ ⊚ Sun light ⊚⊚ 0 0 Example 2 R.T. ⊚ ⊚ ⊚ ⊚ 40° C. ⊚ ⊚ 0 0  4° C. ⊚ ⊚ ⊚ ⊚ Sun light ⊚ ⊚0 0 Example 3 R.T. ⊚ ⊚ ⊚ ⊚ 40° C. ⊚ ⊚ 0 0  4° C. ⊚ ⊚ ⊚ ⊚ Sun light ⊚ ⊚ 00 Example 4 R.T. ⊚ ⊚ ⊚ ⊚ 40° C. ⊚ ⊚ 0 0  4° C. ⊚ ⊚ ⊚ ⊚ Sun light ⊚ ⊚ 0 0Example 6 R.T. ⊚ ⊚ ⊚ ⊚ 40° C. ⊚ ⊚ 0 0  4° C. ⊚ ⊚ ⊚ ⊚ Sun light ⊚ ⊚ 0 0Comparative R.T. ⊚ ⊚ ⊚ 0 Example 2 40° C. ⊚ 0 0 ▾  4° C. ⊚ 0 ▾ ⋆ Sunlight 0 ▾ ⋆ ⋆ Comparative R.T. ⊚ ⊚ 0 ▾ Example 3 40° C. ⊚ 0 ▾ ⋆  4° C. ⊚0 ▾ ⋆ Sun light 0 ▾ ⋆ ⋆ Comparative R.T. ⊚ ⊚ 0 ▾ Example 4 40° C. ⊚ 0 ▾⋆  4° C. ⊚ 0 ▾ ⋆ Sun light 0 ▾ ⋆ ⋆ Comparative R.T. 0 ▾ ⋆ ⋆ Example 540° C. 0 ▾ ⋆ ⋆  4° C. 0 ▾ ▾ ⋆ Sun light ▾ ⋆ ⋆ ⋆R.T.: room temperature⊚ no change,0: slight discoloration,▾: remarkable discoloration, slight phase separation, or suspension,⋆ complete discoloration, phase separation, or precipitation

As shown in Table 6, with respect to the compositions of ComparativeExamples 3 and 5 in which hydroxypropyl beta cyclodextrin orpolyethyleneglycol was not used as a solvent, crystal precipitation atlow temperature occurred (see Comparative Examples 3 and 5) and severeoff-flavor by disulfur was generated (see Comparative Example 5). Thecomposition of Comparative Example 4 containing no phellodendron extractand alteromonas ferment extract exhibited severe browning phenomenonupon exposure to sun light, as compared to the composition of Example 4containing the phellodendron extract and the alteromonas fermentextract. It is assumed that this is caused by UV shielding effect of thephellodendron extract. It had been well known that phellodendron plantexhibits antifungal, antiinflammatory, and antivirus effects in skin.However, there had been no reports that the phellodendron plant canprevent discoloration and off-flavor by exposure to UV light. Thepresent inventors found that the phellodendron plant can prevent phaseinstability of effective components in cosmetic compositions by sunlight and the like.

EXPERIMENTAL EXAMPLE 5 Long-Term Storage Stability

To evaluate the long-term storage stability of the resveratrol used asthe effective component in the stabilized compositions of the presentinvention, the compositions of Example 4 and Comparative Example 5 werestored in a water bath, which had been set to room temperature (R.T.),40° C., and 4° C., for 3 months, and HPLC analysis for the resveratrolwas performed. The test results of long-term storage stability for theresveratrol are summarized in Table 7 below. TABLE 7 Long-term storagestability of resveratrol Example 4 Comparative Example 5 Section R.T.40° C. 4° C. R.T. 40° C. 4° C. Initial 100 100 100 100 100 100 1 month98.01 97.50 98.50 85.20 80.46 75.46 3 months 95.20 94.30 96.40 76.4570.25 65.25

HPLC analysis conditions for the long-term storage stability test ofresveratrol were as follows:

1) Analysis column: Inertsil C8, 120A, Sum, 4.6×150 mm (GL science)

2) Mobile phase: mobile phase 1; 25 mM NaH₂PO₄, mobile phase 2;acetonitrile TABLE 8 HPLC analysis Time (min) Mobile phase 1 Mobilephase 2 Speed (ml/min) 0 100 0 1.0 10.0 80 20 1.5 15.0 50 50 1.5 20.0100 0 1.0 30.0 100 0 1.0

3) Column oven temperature: room temperature

4) Wavelength: 310 nm

5) Load amount: 10 μl

As shown in Table 8, the composition of Comparative Example 5, in whichresveratrol was dissolved in a common solvent, exhibited poor long-termstorage stability due to crystal precipitation at low temperature andphase instability at high temperature. On the other hand, thecomposition of Example 4, in which resveratrol was encapsulated withhydroxypropyl beta cyclodextrin, exhibited excellent long-term storagestability.

The test results for long-term storage stability of alpha-lipoic acidare summarized in Table 9 below. TABLE 9 Long-term storage stability ofalpha-lipoic acid Example 4 Comparative Example 5 Section R.T. 40° C. 4°C. R.T. 40° C. 4° C. Initial 100 100 100 100 100 100 1 month 97.10 96.4098.10 83.90 80.46 85.60 3 months 94.50 94.60 98.50 76.20 70.50 80.20

HPLC analysis conditions for the long-term storage stability test ofalpha-lipoic acid were as follows:

1) Analysis column: Novapak C18, 120A, Sum, 4.6×150 mm (GL science)

2) Mobile phase: mobile phase 1; 0.1% trifluoroacetic acid (pH 2.4),mobile phase 2; acetonitrile TABLE 10 HPLC analysis Speed Time (min)Mobile phase 1 Mobile phase 2 (ml/min) 0 100 0 1.0 3.0 100 0 1.0 8.0 5050 1.0 13.0 50 50 1.0 20.0 100 0 1.0 25.0 100 0 1.0

3) Column oven temperature: room temperature

4) Wavelength: 333 nm

5) Load amount: 20 μl

As shown in Table 9, the composition of Comparative Example 5, in whichalpha-lipoic acid was dissolved in a common solvent, exhibited poorlong-term storage stability due to phase instability with increase oftemperature. On the other hand, the composition of Example 4, in whichalpha-lipoic acid was encapsulated with hydroxypropyl beta cyclodextrin,exhibited excellent long-term storage stability.

EXPERIMENTAL EXAMPLE 6 Skin Irritation Test

To evaluate skin safety, patch test in skins of healthy persons wasperformed using the compositions of Examples 4 and 5 and ComparativeExamples 4 and 5 and the results are summarized in Table 11 below. Testmethod and evaluation method are as described in the above ExperimentalExample 4. TABLE 11 Test results of skin irritation Irritation Degree ofSample score irritation Example 4 1.3 Minimal Example 5 2.1 MildComparative 3.0 Normal Example 4 Comparative 4.5 Severe Example 5

As shown in Table 11, the compositions of Examples 4 and 5, in which aprimary stabilizer of the present invention was used, exhibited moreexcellent skin safety. The composition of Example 5, in which thephellodenderon extract was used as a stabilizer, exhibited an irritationrelief effect by excellent antiinflammatory activity, as compared to thecomposition of Comparative Example 4. In particular, the composition ofExample 4, in which both the phellodendron extract and the alteromonasextract were used, exhibited more excellent skin irritation reliefeffect.

EXPERIMENTAL EXAMPLE 7 Long-Term Effect Test of Antioxidative Activity

To evaluate the long-term effects of the stabilized compositions of thepresent invention, a free radical scavenging activity test for thelong-term effect of antioxidative activity of resveratrol andalpha-lipoic acid was performed and the results are summarized in Table12 below.

The free radical scavenging activity test was performed as follows. Eachsample of the compositions of Example 4, Comparative Examples 4 and 5was collected at an initial time of preparation, 1 month and 3 monthsafter the preparation. The collected sample was dissolved in 10 mg/ml ofethanol and, if necessary, diluted with ethanol, to obtain a samplesolution. 1 ml of 0.1 mM solution of deep blue-coloredDPPH(1,1-diphenyl-2-picryl-hydrazyl) radical in methanol was added to 1ml of the sample solution, vigorously stirred for about 10 minutes, andincubated at 37° C. for 30 minutes. Then, the absorbance (OD_(exp)) ofthe resultant solution was measured at 516 nm by usingspectrophotometer. Here, the degree of reduction of the absorbance wasmeasured because the sample solution was changed from blue color tocolorless as DPPH radical was scavenged. In addition, the samplesolution containing no DPPH solution was used as control sample and theabsorbance (OD_(con)) was measured. The absorbance (OD_(std)) ofstandard sample containing only DPPH solution was also measured.

Free radical scavenging rates were calculated from the above absorbancevalues. The concentrations for 50% free radical scavenging, i.e., IC₅₀were calculated. The calculation method is as described in the aboveExperimental Example 2. As the value of IC₅₀ decreases, a free radicalscavenging activity is excellent. TABLE 12 Free radical scavengingactivity IC₅₀ (μg/ml) Sample Initial 1 month 3 months Example 4 35 42 50Comparative Example 4 40 80 >100 Comparative 48 >100 >1000 Example 5

As shown in Table 12, the composition of Example 4, in which theresveratrol and the alph-lipoic acid with excellent antioxidativeactivity but relatively poor long-term effect of antioxidative activitywere stabilized by the primary stabilizer and the secondary stabilizer,exhibited an excellent long-term effect, as compared to the compositionsof Comparative Examples 4 and 5.

EXPERIMENTAL EXAMPLE 8 Whitening Activity Test Based on Proliferation ofSkin Melanocytes

To evaluate whitening activity, inhibitory activity of the compositionsof Examples 7 and Comparative Examples 6-8 against melanin production inmelanocytes of healthy persons were measured and the results aresummarized in Table 13 below. TABLE 13 Inhibitory activity of melaninproduction according to the content of the water-soluble whiteningcomposition Sample (concentration: Inhibitory activity of melanin 0.5%)production (%) Example 7 75.05 Comparative Example 6 45.26 ComparativeExample 7 11.20 Comparative Example 8 26.58

EXAMPLE 8 Preparation of Skin Toner

According to the composition ratios presented in Table 14 below, twocomponents of a water part and five components of an alcohol part wererespectively mixed with easy-mixer at room temperature. The alcohol partwas gradually added to the water part and solubilized. After theaddition of the alcohol part was completed, the reaction solution wasfurther stirred for 10-20 minutes to obtain a skin toner. TABLE 14 Skintoner Section Component Content Content Water part Purified water To 100To 100 Stabilized composition 6.0 — (Example 4) Alcohol part Ethanol 6.06.0 Hydroxy castor oil 1.0 1.0 POE (60) Flavor Optimal OptimalPreservative Optimal Optimal Dimethicone oil 0.2 0.2

EXAMPLE 9 Preparation of Lotion

According to the composition ratios presented in Table 15 below, a waterpart containing five components and an oil part containing ninecomponents were respectively heated at 75-80° C. until each componentwas completely dissolved. Then, the oil part was gradually added to thewater part and primarily emulsified with homo-mixer at 75-80° C. at 3,000 rpm for 5 minutes. During the primary emulsification, a neutralizerwas added to the reaction mixture. After the primary emulsification wasterminated, the resultant primary emulsion was cooled. Then, a flavorused as an additive was added at 50-55° C., secondarily emulsified withhomo-mixer at 3,000 rpm for 3 minutes, and cooled to 27-30° C., tothereby obtain a lotion. TABLE 15 Lotion Section Component Content Waterpart Purified water To 100 Stabilized composition (Example 4) 30Preservative Optimal Carbomer 0.20 Xanthan gum 0.05 Oil part Cetostearylalcohol 1.0 Self-emulsifiable glycerine 1.0 monostearate Sorbitanmonostearate¹⁾ 0.5 Glycerylmonostearate (POE40)²⁾ 1.0 Liquid paraffin4.0 Squalane 4.0 Cetyloctanoate 6.0 Vasselin 0.3 Beeswax 0.3 NeutralizerTriethanolamine 0.2 Additive Flavor Optimal¹⁾Ar-60 (ICI, USA)²⁾My-52 (ICI, USA)

EXAMPLE 10 Preparation of Cream

According to the composition ratios presented in Table 16 below, a waterpart containing five components and an oil part containing ninecomponents were respectively heated at 75-80° C. until each componentwas completely dissolved. Then, the oil part was gradually added to thewater part and primarily emulsified with homo-mixer at 75-80° C. at3,000 rpm for 5 minutes. During the primary emulsification, aneutralizer was added to the reaction mixture. After the primaryemulsification was terminated, the resultant primary emulsion wascooled. Then, a flavor used as an additive was added at 50-55° C.,secondarily emulsified with homo-mixer at 3,000 rpm for 3 minutes, andcooled to 27-30° C., to thereby obtain a cream. TABLE 16 Cream SectionComponent Content Water part Purified water To 100 Stabilizedcomposition (Example 4) 40 Preservative Optimal Carbomer 0.30 Xanthangum 0.05 Oil part Cetostearyl alcohol 1.5 Self-emulsifiable glycerine2.0 monostearate Sorbitan monostearate¹⁾ 1.0 Glycerylmonostearate(POE40)²⁾ 1.5 Liquid paraffin 6.0 Squalane 5.0 Cetyloctanoate 5.0Vasselin 0.5 Beeswax 1.6 Neutralizer Triethanolamine 0.3 Additive FlavorOptimal¹⁾Ar-60 (ICI, USA)²⁾My-52 (ICI, USA)

EXAMPLE 11 Preparation of Essence

According to the composition ratios presented in Table 17 below, a waterpart containing five components and an oil part containing fivecomponents were respectively heated at 75-80° C. until each componentwas completely dissolved. Then, the oil part was gradually added to thewater part and primarily emulsified with homo-mixer at 75-80° C. at3,000 rpm for 5 minutes. During the emulsification, a neutralizer wasadded to the reaction mixture. After the primary emulsification wasterminated, the resultant primary emulsion was cooled. Then, a flavorused as an additive was added at 5 0-55° C., secondarily emulsified withhomo-mixer at 3,000 rpm for 3 minutes, and cooled to 27-30° C., tothereby obtain an essence. TABLE 17 Essence Section Component ContentWater part Purified water To 100 Stabilized composition (Example 4) 45Preservative Optimal Carbomer 0.40 Xanthan gum 0.10 Oil part Cetostearylalcohol 1.20 Self-emulsifiable glycerine 1.00 monostearate Hydroxycastor oil POE (60) 1.00 Cetyloctanoate 5.00 Cyclomethicone 3.00Neutralizer Triethanolamine 0.40 Additive Flavor Optimal

EXPERIMENTAL EXAMPLE 9 Whitening Activity Test in Human Bodies

The back portions of 20 healthy males/females (age 20 or more) wereartificially blackened by a UV irradiator. Then, the appropriate amountof each of a test product and a control product was applied to the backportions of the test subjects, once a day for 6 weeks. The degree ofwhite and black of skin was measured using chromameter (MINOLTA CR-300,Japan). Skin whitening activity was evaluated by measuring L, a, and bvalues of pigmented skin spots. In detail, skin whitening activity wasevaluated by measuring L, a, and b values of the pigmented skin spots ofthe test subjects using Chromameter CR-300 at 0 weeks (0W), 1 week (1W),2 weeks (2W), 3 weeks (3W), 4 weeks (4W), and 6 weeks (6W) afterexposure to UV light (three times). Chromameter is an apparatus thatrepresents colors using digital codes expressed by three parameters andis widely used to measure a skin color in the cosmetic industry[Muizzuddin N., Marenus K., Maes D., Smith W. P., Use of a chromameterin assessing the efficacy of anti-irritants and tanning accelerators,Journal of the society of Cosmetic Chemists, 1990; 41:369-378). Inaddition, the independent t-test (hypothesized mean difference: 5%) wasperformed to determine if there is a significance difference between thetest product and the control product. TABLE 18 Whitening activity inhuman bodies Section Comparative Example 9 Example 8 0W 65.00 64.66 1W64.71 65.20 2W 65.84 66.13 3W 66.25 66.73 4W 66.34 66.68 5W 66.49 66.746W 65.64 66.34 P (T <= t) 0.028806 Result value 0.92 1.52

As shown in Table 18, the composition of Example 8 according to thepresent invention exhibited excellent whitening activity at astatistically significant level (p<0.05), as compared to the compositionof Comparative Example 9.

INDUSTRIAL APPLICABILITY

As apparent from the above description, a whitening and antioxidativecosmetic composition containing resveratrol of the present invention canprovide excellent whitening and antioxidative effect even when used insmall quantity. Furthermore, the cosmetic composition can stably containthe resveratrol. Even when it is stored for a long term, the cosmeticcomposition sustains excellent whitening and antioxidative effectwithout exhibiting problems such as discoloration, off-flavor, andcrystal precipitation. In addition, the cosmetic composition exhibits askin irritation relief effect.

While the present invention has been particularly shown and describedwith reference to exemplary embodiments thereof, it will be understoodby those of ordinary skill in the art that various changes in form anddetails may be made therein without departing from the spirit and scopeof the present invention as defined by the following claims.

1. A whitening and antioxidative cosmetic composition comprisingresveratrol.
 2. The whitening and antioxidative cosmetic composition ofclaim 1, wherein the resveratrol is used in an amount of 0.001-10.0 wt%, based on the total weight of the cosmetic composition.
 3. Thewhitening and antioxidative cosmetic composition of claim 1, furthercomprising a primary stabilizer selected from the group consisting ofcyclodextrin, polyethyleneglycol, and a mixture thereof.
 4. Thewhitening and antioxidative cosmetic composition of claim 3, wherein theprimary stabilizer is used in an amount of 0.1-50 wt %, based on thetotal weight of the cosmetic composition.
 5. The whitening andantioxidative cosmetic composition of claim 3, further comprising asecondary stabilizer selected from the group consisting of analpha-lipoic acid, a water-soluble whitening composition, aphellodendron extract, an alteromonas ferment extract, and a mixturethereof.
 6. The whitening and antioxidative cosmetic composition ofclaim 5, wherein the secondary stabilizer is used in an amount of0.01-15.0 wt %, based on the total weight of the cosmetic composition.7. The whitening and antioxidative cosmetic composition of claim 5,wherein the water-soluble whitening composition is selected from thegroup consisting of a citrus unshiu peel extract, anaminoethylphosphinic acid, Waltheria indica extract, Guava Phenone, anaqueous hydrolyzed yeast solution, and a mixture thereof.
 8. A methodfor preparing a whitening and antioxidative cosmetic composition,comprising: (a) mixing a solvent and a primary stabilizer; (b) addingresveratrol to the mixture of step (a); and (c) adding a secondarystabilizer selected from the group consisting of an alpha-lipoic acid, awater-soluble whitening composition, a phellodendron extract, analteromonas ferment extract, and a mixture thereof, to the mixture ofstep (b).